The capacity to reprogram bacteria to carry out useful tasks builds upon existing genetic circuit elements that can be repurposed. Two challenges to the design of synthetic bio-circuits include coordination of expression from multiple genes,and integration of metabolite feedback for expression control. Engineered trans-acting RNAs are flexible tools for rapid and specific control of gene expression. Naturally occurring bacterial small regulatory RNAs (sRNAs) can simultaneously target multiple mRNA transcripts via specific base-pairing, and can either stabilize or destabilize target transcripts. The importance and activity of these native small nucleic acids have been established, and it is time for their development for applications. The goal of the research is to create a ligand-responsive riboswitch-sRNA fusion molecule for improved fermentation cultures that produce biofuels and medicines.
The Engineering of multi-target sRNA will enable coordinate and discoordinate mRNA stability/ translation control for metabolic engineering. Riboswitches integrated for control of RS-sRNAs will create a platform for user-specified targeting of mRNAs and integration with riboswitchbound metabolite feedback. Riboswitches integrated for stability control of sRNA will probe sRNA structural dynamics and activity, and will inform future design considerations for engineered sRNAs in their biological context.
The proposed work will create fundamental tools for synthetic circuit development from the sRNAs, advance biochemical engineering by further bridging the disciplines of engineering and genetics, and enable us to train the next generation of biochemical engineers. The construction and analysis of modified sRNA genes can be parsed into smaller project subunits amenable to inclusion of undergraduates and summer students.
